CellularandMolecularBiologyoftheNeuron

gy
PartII

YanZhang

Cellular Information Transfer

Directly contacted cells: cellular junctions and recognition

Non-contacted cells: chemical communication

Chemicals regulating
g
g cellular life activities are called signaling
g
g molecules.

General steps of chemical information transduction

Certain cells release signaling molecules

Signaling molecules reach target cells

by diffusion or blood circulation
Bind to receptors
p
of target
g cells specifically
p
y

Receptors transduce signals and activate intracellular messenger system

Target cells trigger biological effects

Extracellular signaling molecules: ligand or first messenger

Chemicals secreted by cells to regulate life activities of target cells
Low dosage
dosage, short lifespan
lifespan, high efficiency and high specificity
Types: autocrine, paracrine and endocrine

chemicals: proteins and polypeptides, amino acids and their derivatives,

steroid hormones, fatty acid derivatives and gas
solubleness: water-soluble, liposoluble and gas

Chemicals transmitting intracellular

information after first messenger stimulation

Secondary messengers
Small molecules transmitting information intracellularly, such as Ca2+, DAG, IP3, Cer,
cAMP, cGMP, arachidonic acid and its metabolites.

Third messengers
Molecules transmitting information through the nucleus. They are nucleoproteins
binding to specific sequences of target genes and are called DNA binding proteins as
well.
6

Chemical properties

Inorganic ions: such as Ca2+

Fat derivatives: such as DAG and Cer

Nucleotides: such as cAMP and cGMP

Sugar derivatives: such as IP3

Signaling
g
gp
proteins
7

CNS neurotransmitters
CNSneurotransmitters
1.:
Concept:chemicalsinvolvedinsynaptictransmissioninthecentralnervoussystem(CNS)

:
Qualifications:
Q
lifi ti

Producibleinpresynapticneuronscontainingits raw materialsandenzymicsystems

Storedinsynapticvesiclesandreleasedtothesynapticcleftswhenimpulsionarrives

Bindtopostsynapticreceptorstotriggerspecificphysiologicaleffects

Abletobeinactivatedbyenzymesorotherapproachessuchasreuptaking

y
y
g
Itseffectscanbeenhancedbyneurotransmittermimicsandblockedbyantagonists
8

2. Typesofneurotransmitters

TypesMembers

Cholines
AcetylcholineAch

Monoamines

NEE
Catecholaminedopamine,norepinephrine andepinephrine

5HT
Indoleamine:5HT

GABA
GABA
Aminoacids glutamic acid,asparticacid,glycine andGABA

ADH
Peptides:
Peptides:hypothalamusregulatorypeptide,ADH,oxytocin,opioid,braingut
hypothalamus regulatory peptide ADH oxytocin opioid brain gut
peptide,atrial natriuretic peptide.

ATP
Purines:
Purines:adenosine,ATP
adenosine ATP

NOCO
Gas:NO,CO

PG
Lipids:PG

3. Receptor doctrine
1

Conceptmacromolecular proteins set in cellular membrane, recognizes and binds

to specific transmitters, triggers physiological effects and alters the cell membrane
permeabilit of certain ions.
permeability
ions

ligand
ga d

Agonist
Antagonist

2
Binding specificity of receptors and ligands: specificity, saturability, reversibility

3Sorted
3
S t db
by:
Position pre- and post-synaptic

coupled to ion channels

Biological effects G
activate G proteins and protein kinase receptors

cholinergic receptors
Binding transmitters adrenergic receptors
presynaptic receptors

5-HT

CNS transmitter receptors:
5-HT receptors, amino acid receptors

 Major transmitterreceptor systems

TransmitterReceptorSecondmessengerAntagonistChanneleffectMajordistribution

N1

Muscle-type nicotinic receptor

Decamethonium

N2

Neuron-type nicotinic receptor

Hexamethonium

ACh

M1
M2

IP3/DG

M3

cAMP

M4

IP3/DG

cAMP

( Cardiac)

()

Atropine

Na+

PNS
Na+and
other
Allautonomicnervepreganglionic fibers,most
small ions
parasympatheticpostganglionicfibers,afew
sympatheticpostganglionicfibers,skeletal
musclefibers
l fib

Ca2+
K+

CNS:
Spinalcordanteriorhornmotorneurons,
posteriorventralthalamusspecificsensory
projectionneurons,brainstemreticular
g
g y
,
,
structureascendingexcitingsystem,striatum,
limbicsystem

TransmitterReceptorSecondmessengerAntagonistChanneleffectMajordistribution

1 IP3/DG
2

Phentolamine

cAMP

small intestine

Cardiac

Phentolamine

Presynaptic membrane

NE

cAMP
)

K+
K+
Ca2+

Yohimbine

Practolol

At l l
Atenolol

2
D1D5

Dopamine

5-HT

D 2D 3D 4

5-HT1
5-HT2

butoxamine

cAMP
cAMP
cAMP
IP3/DG

K+
C 2+
Ca

PNS:
Most parasympathetic postganglionic fibers
Mostparasympatheticpostganglionicfibers
CNS:
Lowerbrainstemandascendingfibers
projectingtothecortex,limbicforebrain,
hypothalamusanddescendingfibers
projectingtoposterior/lateral/anteriorhorn
ofthespinalcord

Substantia nigrastriatum,tubero
infundibular system,midbrainlimbicsystem
system, midbrain limbic system

K+

Insideraphe

I id
h nucleiandascendingfibers
l i d
di fib

K+

projectingtostriatum,hypothalamusetc.and
descendingfiberstoto posterior/lateral/
anteriorhornofthespinalcord

45

()

Motorendplate microscopy(AuCl3 staining)


Motor endplate ultrastructure


Motor endplate SEM image

Neuronal soma
NeuronalsomaSynapticknob
Synaptic knob

Neuronaalsomaandsuperrficialsyn
naptickn
nobsbySSEM

Themechanismsignalmoleculesactontargetingcells

Receptors
1 Concept
1.
1.

M
Macromoleculesincellularmembranesorcytoplasmthatrecognizeandbind
l l i
ll l
b
t l
th t
i
d bi d
bioactivemolecules(hormones,neurotransmitters,toxinsanddrugs).

Majorlyproteinsandminorly glycolipids

DNA
DNA

Receptorsaremajorlylocatedinthemembrane(membranereceptors
Receptorsaremajorlylocatedinthemembrane(membranereceptors,majorly
,majorly
integralglycoproteins
integral
glycoproteins)and
)andminorly
minorly inthecytoplasm(intracellularreceptors
inthecytoplasm(intracellularreceptors,all
,all
are DNA binding proteins)
areDNAbindingproteins).

ligand

ligand

Bioactivemoleculesbindingtoreceptors.Theyrecarriersofinformationand
arealsocalledfirstmessengers.

2 Characteristicsofreceptors
Characteristics of receptors

Specificity
p
y

Characteristics of receptors
p

Saturability

Reversibility

High affinity

Specific action mode

3 Classificationofmembranereceptors
G- GPCR
GPCR
epinephrine
epinephrine
glycogen
glycogen
EnzymeEnzyme-linked receptors:
receptors: growth factors
Ion channelreceptors
channelreceptors
Acetylcholine
G protein coupled receptors (GPCR)

G protein

Ligand

Activated G protein
Enzymes or ion channels

Activated enzymes or ion channels

Ligand

Enzyme-linked receptors
Non-activated catalytic site

Activated catalytic site

Ions
Ligand

Ion channels

Transmembranesignaltransduction

Extracellular signal molecules bind to membrane receptors

and transduce information into the cytoplasm or nucleus,
regulating functions of targeting cells.

G protein: an peripheral protein located in the cytoplasm and

g to GTP/GDP,, transducing
g extracellular information to
binding
intracellular information and named after its close relationship
with GDP/GTP.

Heterotrimer composed of // subunits

Small molecule G protein

G GPCR system

GG GTP
GTP
GTP
GTP

GDP
GDP

G protein: GTPGTP-binding regulatory protein. Binds to GTP/GDP.

Composed of // subunits.

Two conformations

GDP
Non-activated

GTP

Activated

Widely distributed and involved in regulations of material metabolism and gene transcription.

Common G protein types

Type

subunit

Function
Activate adenylate cyclase

Inhibit adenylate cyclase

Activate p
phosphatidylinositol
p
y
p
phospholipase
p
p
C

Major
j Gp
protein in brain and regulates
g
ion channels

Activate vision

G7
-
GPCR: single-chain transmembrane peptide, with 7
transmembrane helixes shuttling in the membrane

() G
Major path a s of GPCR signal transd ction
MajorpathwaysofGPCRsignaltransduction
GAC
R
ReceptorGproteinAC(adenylate
t G
t i AC ( d l t cyclase)pathway
l ) th
cAMPAprotein kinase A,PKA

Charactered bv cAMP concentrationalterationsintargetcellsandPKA

activation,majorwayforhormonestoregulatemateral metabolism

Function: regulations of material metabolism and gene transcription

cAMP signal transduction pathway (PKA pathway)

Ligand
g

M b
Membrane
receptor
t

Gs protein
((Transducer))

Membrane
Second messenger
g enzyme
y
target

ACadenylyl cyclase
PKAprotein kinase A

Effector

Biological effects

Hormones(likeepinephrine)

Extracellular

AC

Celllular
membrane

Ligand bindstoreceptors

Intracellular

Gprotein

GTP/GDPG

Phosphodiesterase
p
Nonactivated
PKA

Exchange GTP/GDP (G protein activation)

ExchangeGTP/GDP(Gproteinactivation)

Activated
PKA

AC
Bi d d i
BindandactivateAC(adenylate
AC ( d l
cyclase)
l )

Phosphorylase
b kinase
bkinase
(notactivated)

Phosphorylase
b kinase
bkinase
GeneratecAMP (secondmessenger)
(activated)

cAMP

Phosphorylase bkinase phosphatase

PKA
Phosphorylase b

Phosphorylase a

Phosphorylase a
h h
phosphatase

Activte PKA

Correspondingeffects
Glycogen glucose1phosphate

PLC signal
g
transduction p
pathwayPKC
y

Ligand
Gp protein
Membrane
receptors

integral
g

Biological effects
PLC phospholipase C
IP3 Inositol triphosphate
DAG diacylglycerol
PKCprotein kinase C
C M
CaMcalmodulin
l d li


EnzymeEnzyme
-linked receptor mediated signal transduction system
G
G
Enzyme linkedreceptorsaredistinctivefromGPCRsinmolecularstructuresand
Enzymelinked
receptors are distinctive from GPCRs in molecular structures and
characteristics,whosecytoplasmicsideactsasanenzymeorbindsdirectlyand
activatescytoplasmicenzymeswithoutGproteins.

6typesofknownenzymelinkedreceptors: receptortyrosinekinase receptor

guanylatecyclase receptortyrosinephospholipaseReceptorserine/threonine
kinaseTyrosinekinaselinkedreceptorHistidinekinaselinkedreceptorrelated
tothechemotaxisofbacteria

Followingismainlyonreceptortyrosinekinase
ll
l
k
andreceptorguanylatecyclase.
d
l
l


Receptortyrosinekinase

receptorproteintyrosine
kinasesRPTKs
RPTKs

Receptorproteintyrosinekinasesarethelargest
typeofenzymelinkedreceptors.RPTKsaremade
of3compartments:extracellulardomain,single
f3
ll l d
i i l
passhydrophobichelixandintracellulardomain.
TheextracellulardomainofRPTKsisligandbinding
region, and the ligand is soluble or membrane
region,andtheligandissolubleormembrane
boundpeptideorproteinhormonesincluding
insulinandvariousgrowthfactors.Theintracellular
domainisthecatalyticsiteoftyrosineprotein
ki
kinaseandownsautophosphorylationsites.
d
h h l i
i

EGF

The ligand (such as EGF) binds to the receptor in the extracellular domain and
Theligand(suchasEGF)bindstothereceptorintheextracellulardomainand
triggersconformationalchanges,leadingtoreceptordimerization andformation
ofahomodimer orheterodimer andcausingautophosphorylation inthe
intracellulartyrosineresidues,thusactivatingtheirtyrosineproteinkinase
ll l
d
h
h
k
activity.
Theintracellulardomainsofthedimer formasignaltransductioncomplex.

Receptor TPK-Ras-MAPK pathway

Growth factor
Membrane receptor

Receptor
dimerization

Receptor
TPK activation

Cam Kinase

Transcription factor motification

Biological effects

Biological effects

Gene expression regulation

JAK /STAT pathway

JAKs/STAT
th
Growth factor
Membrane receptor Receptor
dimerization

Cytosolic JAKs
activation

STAT
activation

Gene
expression
regulation

Soluble receptors

Atrial natriuretic factor

Receptor guanylate cyclase
Membrane receptors
Effector protein
phosphorylation

Bi l i l effects
Biological
ff t

Hormone

Receptor

G protein

Enzyme

Second messenger

Protein kinase

Enzyme or other functional proteins

Bi l i l effects
Biological
ff t


Ion channel receptor
receptor-mediated signal transduction

(gating)
Ionchannelreceptorsareatypeofreceptorsalsofunctioningasionchannels.The
openingand
d closingofachanneliscalledgating.
l
f h
l
ll d
(1)(2)
( )
(3)3,
,

Ionchannelsareclassifiedinto3typesaccordingtogatingmechanisms:(1)chemically
gated channels (2)voltage gated channels (3) mechanically gated channels These 3 types
gatedchannels(2)voltagegatedchannels(3)mechanicallygatedchannels.These3types
ofchannelproteinsenabledifferentcellstoreacttoenvironmentalstimulationsand
conducttransmembranesignaltransductions.

Ionchannelreceptor
Ion
channel receptormediated
mediatedsignaltransductiondoesn
signal transduction doesnttneedtogenerateother
need to generate other
intracellularmessengersandarequickandsitelimited.

 Chemicallygatedionchannel
:

Chemicallygatedionchannelsarealsocalledligandgatedionchannelsandarenamed
yg
g
g
bycorrespondingtransmitterssuchasacetylcholine receptorchannels,glutamine
receptorchannelsandaspartatereceptorchannels.
N2Ach4

5
2

4 5 A h
45Ach

N2Achreceptorcationchannelsare
madeof5subunitsof4types,forming
a2
pentagonshapedchannel
p
g
p
.
Everysubunitpassesthroughthe
membrane4times;Achbindingsiteis
locatedinthesubunitanditsbinding
forces the channel to open and facilited
forcesthechanneltoopenandfacilited
diffusionoftheionshelpstofinishthe
signaltransductionprocess.

 Voltagegatedionchannels
:
K+Na+Ca2+Cl4
Voltagegatedionchannelsarealsocalledvoltagedependentorvoltagesensitiveionchannels,and
theyopenandclosedependingonthechangesofthemembranepotentialandarenamedafterthe
ionspassingthroughmosteasilysuchasK+Na+Ca2+Cl channelsandtheyarefurtherdivided
intoseveralsubtypes.

Theirmolecularstructureissimilartochemicalchannelsbuttherearesomedomainssensitiveto
transmembranepotentialchangesandtriggersthechangesofthewholechannel.

Mechanicallygatedchannels
yg
:

Mechanicallygatedchannelsarealsocalledmechanosensitivechannelsandtheyareatypeof
channelssensingvariationofsuperficialcellularmembranestressesandthustransduceextracellular
mechanicalsignalstointracellularsignals.Theyareclassifiedtoionselectiveandnonselective
channelsbypermeabilityandtostretchactivatedandstretchinactivatedchannelsbyfunction.

Cytosolic
y
and intranuclear receptors
p
and their signal
g
transduction
Intranuclear receptors

Ligands enter cells

Bind to intranuclear
hormone response
elements

Ligand-receptor
complex

Ligand-receptor
dimerization
Gene expression
regulation

Cytosol
y
Receptor-ligand complex

ReceptorNucleus
Transcription
i iti ti complex
initiation
l

Activated receptorligand complex

Gene
Translation

Biological effects

Proteins


Crosstalk of cellular signal transduction pathways

Single piece of information acts on multiple pathways

Insulin receptor substrate

PKC pathway

Receptor TPK activation

PKT pathway

Multiple pathwayy signal moleculess

Insulin

Components of a single pathway regulates another pathway

Thyrotropin-releasing hormone

PKC pathway

PKA pathway

Pa
athway re
egulation
n

Adenylate cyclase

Distinctive pathways regulates the same protein

Gene expression regulation

Efffector pro
oteins of multiple pathwayss

P
PKC
pathw
way

P
PKA
pathw
way
CaM kinase

Neuroscience

return

 LTP
LTP

Requirements of LTP generation

Activation of synapse
Strongg depolarization
p
of p
postsynaptic
y p cell in CA1

Requirements to trigger depolarization of the postsynaptic neuron
AshighasenoughfrequencyEPSPtemporalsummation
As many as enough synapseEPSP
Asmanyasenoughsynapse
EPSPspatialsummation
spatial summation

CA
CA1
1 LTP
LTP
Postsynaptic factors to trigger CA
CA1
1 LTP
postsynatic depolarizaion
activation of NMDA receptors
influx of Ca2+
activation by Ca22+ of several secondmessenger
second messenger systems in the postsynaptic cell

LTPHasaTransientEarlyandaConsolidatedLatePhase
LTP
LTP

EarlyLTP
EarlyLTP
y
onestimulustrainproducesanearly,
p
y,
shorttermphaseofLTP,lasting13hoursNonew
protein synthesis
proteinsynthesis
LTPLTP
1 3
13
LateLTP Fourormoretrainsinduceamore
LateLTP
persistentphaseofLTPthatlastsatleast24hours
q
p
y
requiresnewproteinandRNAsynthesis
LTP4
LTP24RNA
LTP24RNA

Control

(Kandel,ER,JHSchwartzandTMJessell(2000)
PrinciplesofNeuralScience.NewYork:McGrawHill.)
p
f
)

EarlyandlatephasesofLTP
/LTP
/
Theearlyphase
Presynaptic site: increase in probability of
Presynapticsite:increaseinprobabilityof
transmitterrelease,withoutstructure
changes.
:
Thelatephase
p

Presynapticsite:activation,perhapsthegrowth,
Postsynapticsite:
ofadditionalmachineryfortransmitter

release
p
postsynapticdeploarization
y p
p

activationofNMDAreceptors

NMDA
Postsynapticsite:insertionofnewclustersof
influxofcalcium
influx of calcium
receptors

activationofseveralsecondmessenger
systemsbycalcium

Increasethesensitivityandnumberofthe
postsynapticAMPArecepors
AMPA
(Kandel,ER,JHSchwartzandTMJessell(2000)
PrinciplesofNeuralScience.NewYork:McGrawHill.)

MechanismsofEarlyLTP
MechanismsofEarlyLTP
LTP
LTP

Singletraindepolarizationofthemembraneofpostsynapticcell
Si l
i d
l i i
f h
b
f
i
ll

RemovetheblockageofMg2+ toNMDAreceptor
Mg2+ NMDA
NMDA

Ca2+ influxinpostsynapticcellactivationofproteinkinases
influxinpostsynapticcellactivationofproteinkinases
Ca

Ca2+

A ti it h
ActivitychangesofAMPAreceptor
f AMPA
t
AMPA
AMPA

Retrogradesignalgenerator

NO?

Glu
G u isreleasedfrom
isreleasedfrompresynaptic
s e eased o presynaptic
p esy apt c ce
cell

InfluxofNa+
N +
Na

AMPAreceptor
AMPA

NMDAreceptor
NMDA

MechanismsofLateLTP
MechanismsofLateLTP
LTP
LTP

R
Repeatedtrainsdepolarizationofthemembraneofpostsynapticcell
t d t i d
l i ti
f th
b
f
t
ti
ll

RemovetheblockageofMg2+ to
toNMDAreceptor
NMDAreceptor
Mg2+ NMDA
NMDA

Ca2+ influxinpostsynapticcell
Ca

Ca2+
Adenylyl cyclase

Ca2+/calmodulin
Ca2+/
cAMP

cAMP kinase activation

cAMP
cAMP

MAPkinase
MAP
kinase MAP
MAP
Newsynapse

PCREB1
PCREB
Effectors

(tPA
tPA,BDNF)
,BDNF)

CRE

Regulators

(C/EBP
(C/
EBP
)

Molecular mechanisim of learning and memory

cAMP
cAMP
cAMPLTP
cAMP
AMP plays
l
a kkey role
l iin th
the habituation
h bit ti and
d sensitization
iti ti
off the
th gill-withdraw
ill ithd
reflex
fl off
sea slug aplysia.
cAMP signaling
g
gp
pathway
y is crucial in p
protein-synthesis
y
dependent
p
p
phase of LTP

CaMKIILTP
The permanent activation of CaMKII is critical in inducement and maintenance of LTP
PKA
PKA is critical in long-term sensitization

CREB

Impact on long-term memories

Gene expression regulation

cAMPA
CREBCRE
CREBCRE

CREDNAcAMP

 MechanismsofShortTermSensitization

Interneuron

5HT
Goprotein
Go

5 HTR
Sensoryy neuron5
neuron5HTR

Gsprotein
Gs

DAG

PLC

PKC

AC
PKAactivation

ATP

K+ channel
channel

K+ channel closed
K+
K+ conductance
AP
AP
AP duration
APduration

cAMP

PKA
PKA

LtypeCa2+
Channelopen
LCa2+

Vesiclemobilizedtoactivezone

NtypeCa2+
Channelopen
2
NCa2+

Ca2+
Ca2+ influx
C
i fl

T
Transmitter
i
release


Moleculesinvolvedinlong TermSensitization

Interneuron

5HT
MAPK

5 HTR

cAMP

CREB2
CREB

PKA

Sensoryy neuron5
neuron5HTR

PCREB
CREB1
CREEarly
C/EBP
Ubi iti Hydrolase
Ubiquitin
H d l

CAATLate

Transmitter
Release

Persistentactivity
ofPKA
of PKA
PKA
PKA

Growthofnew
synapticconnections
synaptic connections